WO2018002851A1 - Augmentation de la croissance et du rendement des plantes au moyen d'une séquence de thiorédoxine - Google Patents

Augmentation de la croissance et du rendement des plantes au moyen d'une séquence de thiorédoxine Download PDF

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Publication number
WO2018002851A1
WO2018002851A1 PCT/IB2017/053883 IB2017053883W WO2018002851A1 WO 2018002851 A1 WO2018002851 A1 WO 2018002851A1 IB 2017053883 W IB2017053883 W IB 2017053883W WO 2018002851 A1 WO2018002851 A1 WO 2018002851A1
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WIPO (PCT)
Prior art keywords
plant
thioredoxin
protein
expression
sequence
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Application number
PCT/IB2017/053883
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English (en)
Inventor
Benjamin Neil GRAY
Henry D. PRIEST
Original Assignee
Benson Hill Biosystems, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Benson Hill Biosystems, Inc. filed Critical Benson Hill Biosystems, Inc.
Priority to US16/311,905 priority Critical patent/US11371053B2/en
Priority to CN201780041270.9A priority patent/CN109563519A/zh
Priority to EP17743085.7A priority patent/EP3478846A1/fr
Priority to BR112018077178-3A priority patent/BR112018077178A2/pt
Priority to CA3029126A priority patent/CA3029126A1/fr
Publication of WO2018002851A1 publication Critical patent/WO2018002851A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • C12N15/8222—Developmentally regulated expression systems, tissue, organ specific, temporal or spatial regulation
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y108/00—Oxidoreductases acting on sulfur groups as donors (1.8)
    • C12Y108/01—Oxidoreductases acting on sulfur groups as donors (1.8) with NAD+ or NADP+ as acceptor (1.8.1)
    • C12Y108/01008—Protein-disulfide reductase (1.8.1.8), i.e. thioredoxin
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
    • Y02A40/146—Genetically Modified [GMO] plants, e.g. transgenic plants

Definitions

  • the invention is drawn to compositions and methods for increasing plant growth and yield through expression of a thioredoxin gene in a plant.
  • compositions and methods for regulating gene expression in a plant are provided.
  • the methods increase plant growth resulting in higher crop yield.
  • Such methods include increasing the expression of at least one thioredoxin gene in a plant of interest.
  • the invention also encompasses constructs comprising a promoter that drives expression in a plant cell operably linked to a thioredoxin coding sequence.
  • Compositions further comprise plants, plant seeds, plant organs, plant cells, and other plant parts that have increased expression of a thioredoxin sequence.
  • the invention includes methods that can be utilized to increase expression of a thioredoxin gene in a plant.
  • Such thioredoxin gene may be a native sequence or alternatively, may be a sequence that is heterologous to the plant of interest.
  • SEQ ID NO: l comprises SEQ ID NO: l, or encodes a protein selected from the group consisting of SEQ ID NOs:2 and 17-100.
  • the plant of embodiment 12 wherein said plant is from the genus Glycine, Brassica, Medicago, Helianthus, Carthamus, Nicotiana, Solanum, Gossypium, Ipomoea, Manihot, Coffea, Citrus, Theobroma, Camellia, Persea, Ficus, Psidium, Mangifera, Olea, Carica, Anacardium, Macadamia, Prunus, Beta, Populus, or Eucalyptus.
  • the accumulation of harvestable biomass results from plant growth and allocation of photosynthetically fixed carbon to the harvested portion(s) of the plant.
  • Plant growth may be manipulated by modulating the expression of one or more plant genes. This modulation can alter the function of one or more metabolic pathways that contributes to plant growth and accumulation of harvestable biomass.
  • compositions of the invention include constructs comprising the coding sequence set forth in SEQ ID NO: l or encoding a protein selected from the group of SEQ ID NOs:2 and 17-100 or variants thereof, operably linked to a promoter that is functional in a plant cell.
  • promoter is intended to mean a regulatory region of DNA that is capable of driving expression of a sequence in a plant or plant cell. It is recognized that having identified the thioredoxin protein sequences disclosed herein, it is within the state of the art to isolate and identify additional thioredoxin protein sequences and nucleotide sequences encoding thioredoxin protein sequences, for instance through BLAST searches, PCR assays, and the like.
  • a "native" polynucleotide or polypeptide comprises a naturally occurring nucleotide sequence or amino acid sequence, respectively.
  • variants of a particular polynucleotide of the invention will have at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to that particular polynucleotide as determined by sequence alignment programs and parameters as described elsewhere herein.
  • Fragments and variants of the polynucleotides disclosed herein can encode proteins that retain thioredoxin function.
  • Variant amino acid or protein is intended to mean an amino acid or protein derived from the native amino acid or protein by deletion (so-called truncation) of one or more amino acids at the N-terminal and/or C -terminal end of the native protein; deletion and/or addition of one or more amino acids at one or more internal sites in the native protein; or substitution of one or more amino acids at one or more sites in the native protein.
  • Amino acids can be generally categorized as aliphatic, hydroxyl or sulfur/selenium- containing, cyclic, aromatic, basic, or acidic and their amide. Without being limited by theory, conservative amino acid substitutions may be preferable in some cases to non-conservative amino acid substitutions for the generation of variant protein sequences, as conservative substitutions may be more likely than non-conservative substitutions to allow the variant protein to retain its biological activity. Polynucleotides encoding a polypeptide having one or more amino acid substitutions in the sequence are contemplated within the scope of the present invention. Table 1 below provides a listing of examples of amino acids belong to each class.
  • Computer implementations of these mathematical algorithms can be utilized for comparison of sequences to determine sequence identity. Such implementations include, but are not limited to: CLUSTAL in the PC/Gene program (available from Intelligenetics, Mountain View, California); the ALIGN program (Version 2.0) and GAP, BESTFIT, BLAST, FASTA, and TFASTA in the GCG Wisconsin Genetics Software Package, Version 10 (available from Accelrys Inc., 9685 Scranton Road, San Diego, California, USA). Alignments using these programs can be performed using the default parameters.
  • CLUSTAL program is well described by Higgins et al. (1988) Gene 73:237-244 (1988); Higgins et al.
  • Gapped BLAST in BLAST 2.0
  • PSI-BLAST in BLAST 2.0
  • PSI-BLAST in BLAST 2.0
  • Codon optimization is when one or more codons are altered at the nucleic acid level such that the amino acids are not changed but expression in a particular host organism is increased.
  • a number of promoters may be used in the practice of the invention.
  • the polynucleotides encoding a thioredoxin protein of the invention may be expressed from a promoter with a constitutive expression profile.
  • Constitutive promoters include the CaMV 35S promoter (Odell et al. (1985) Nature 313:810-812); rice actin (McElroy et al. (1990) Plant Cell 2: 163-171); ubiquitin (Christensen et al. (1989) Plant Mol. Biol. 12:619-632 and Christensen et al. (1992) Plant Mol. Biol. 18:675-689); pEMU (Last et al. (1991) Theor. Appl. Genet. 81:581-588); MAS (Velten et al. (1984) EMBO J. 3:2723-2730); ALS promoter (U.S. Patent No. 5,659,026), and the
  • Plant terminators are known in the art and include those available from the Ti-plasmid of A. tumefaciens, such as the octopine synthase and nopaline synthase termination regions. See also Guerineau et al. (1991) Mol. Gen. Genet. 262: 141-144; Proudfoot (1991) Cell 64:671-674;
  • nucleotides encoding thioredoxin proteins of the present invention can be used in expression cassettes to transform plants of interest. Transformation protocols as well as protocols for introducing polypeptides or polynucleotide sequences into plants may vary depending on the type of plant or plant cell, i.e., monocot or dicot, targeted for transformation.
  • transformation protocols as well as protocols for introducing polypeptides or polynucleotide sequences into plants may vary depending on the type of plant or plant cell, i.e., monocot or dicot, targeted for transformation.
  • the term “transform” or “transformation” refers to any method used to introduce polypeptides or
  • the cells that have been transformed may be grown into plants in accordance with conventional ways. See, for example, McCormick et al. (1986) Plant Cell Reports 5:81-84.
  • the present invention provides transformed seed (also referred to as "transgenic seed") having a polynucleotide of the invention, for example, an expression cassette of the invention, stably incorporated into their genome.
  • a deactivated Cas9 endonuclease fused to a transcriptional enhancer element is targeted to a genomic location near the transcription start site for a thioredoxin gene of interest, thereby modulating the expression of said thioredoxin gene of interest (Piatek et al. (2015) Plant Biotechnol J 13:578-589).
  • Enhancers include any molecule capable of enhancing gene expression when inserted into the genome of a plant.
  • an enhancer can be inserted in a region of the genome upstream or downstream of a thioredoxin sequence of interest to enhance expression.
  • Enhancers may be exacting, and can be located anywhere within the genome relative to a gene for which expression will be enhanced.
  • an enhancer may be positioned within about 1 Mbp, within about 100 kbp, within about 50kbp, about 30 kbp, about 20 kbp, about 10 kbp, about 5 kbp, about 3kbp, or about lkbp of a coding sequence for which it enhances expression.
  • A. tumefaciens cells harboring thioredoxin plant transformation vectors are used to transform maize (Zea mays cv. B 104) cells suitable for regeneration on tissue culture medium. Following transformation of the maize cells with the relevant plant transformation vectors and regeneration of maize plants, PCR analyses are performed to confirm the presence of the gene(s) of interest in the maize genome.
  • A. tumefaciens cells harboring thioredoxin plant transformation vectors are used to transform rice (Oryza sativa cv. Kitaake) cells suitable for regeneration on tissue culture medium. Following transformation of the rice cells with the relevant plant transformation vectors and regeneration of rice plants, PCR analyses are performed to confirm the presence of the gene(s) of interest in the rice genome.
  • Table 5 Summary of S. viridis greenhouse observations with Tl -generation plants

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  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Botany (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

La présente invention concerne des compositions et des procédés permettant d'améliorer la croissance des plantes. La présente invention concerne des polynucléotides codant pour des protéines de thiorédoxine, des polypeptides comprenant des protéines de thiorédoxine, et des constructions d'expression formulées pour exprimer des gènes d'intérêt dont l'expression peut améliorer les propriétés agronomiques comprenant, mais sans limitation, le rendement des cultures, la tolérance aux agressions biotiques et abiotiques, la vigueur à la levée; l'invention porte également sur des plantes comprenant les polynucléotides, les polypeptides, et les constructions d'expression ainsi que sur des procédés de production de plantes transgéniques.
PCT/IB2017/053883 2016-06-29 2017-06-28 Augmentation de la croissance et du rendement des plantes au moyen d'une séquence de thiorédoxine WO2018002851A1 (fr)

Priority Applications (5)

Application Number Priority Date Filing Date Title
US16/311,905 US11371053B2 (en) 2016-06-29 2017-06-28 Increasing plant growth and yield by expression of an m-type thioredoxin
CN201780041270.9A CN109563519A (zh) 2016-06-29 2017-06-28 使用硫氧还蛋白序列增加植物生长和产量
EP17743085.7A EP3478846A1 (fr) 2016-06-29 2017-06-28 Augmentation de la croissance et du rendement des plantes au moyen d'une séquence de thiorédoxine
BR112018077178-3A BR112018077178A2 (pt) 2016-06-29 2017-06-28 aumento de crescimento e rendimento de planta pelo uso de uma sequência de tiorredoxina
CA3029126A CA3029126A1 (fr) 2016-06-29 2017-06-28 Augmentation de la croissance et du rendement des plantes au moyen d'une sequence de thioredoxine

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201662356120P 2016-06-29 2016-06-29
US62/356,120 2016-06-29

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WO2018002851A1 true WO2018002851A1 (fr) 2018-01-04

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US (1) US11371053B2 (fr)
EP (1) EP3478846A1 (fr)
CN (1) CN109563519A (fr)
BR (1) BR112018077178A2 (fr)
CA (1) CA3029126A1 (fr)
WO (1) WO2018002851A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019239373A1 (fr) * 2018-06-14 2019-12-19 Benson Hill Biosystems, Inc. Augmentation de la croissance et de la productivité des plantes par l'utilisation d'une protéine de la superfamille ring/u-box
WO2022065759A1 (fr) * 2020-09-25 2022-03-31 경상국립대학교산학협력단 Plante transgénique résistante au froid surexprimant la protéine thiorédoxine trx-h2

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024033924A1 (fr) * 2022-08-11 2024-02-15 Futuragene Israel Ltd. Méthodes de sélection et de production de plantes d'eucalyptus résistantes à une perturbation physiologique
CN116640738A (zh) * 2023-05-31 2023-08-25 中国热带农业科学院橡胶研究所 来源于橡胶树的硫氧还蛋白HbTRXy2及其相关生物材料与应用

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